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PMID: 16617147 Published · epublish English Journal Article Research Support, N.I.H., Extramural

Identification and characterization of mitochondrial abasic (AP)-endonuclease in mammalian cells.

Nucleic acids research ·Vol. 34 ·No. 7 ·2006-00-00 ·Pages 2067-76

Chattopadhyay R, Wiederhold L, Szczesny B, Boldogh I, Hazra TK, Izumi T, Mitra S

Abstract

Abasic (AP)-endonuclease (APE) is responsible for repair of AP sites, and single-strand DNA breaks with 3' blocking groups that are generated either spontaneously or during repair of damaged or abnormal bases via the DNA base excision repair (BER) pathway in both nucleus and mitochondria. Mammalian cells express only one nuclear APE, 36 kDa APE1, which is essential for survival. Mammalian mitochondrial (mt) BER enzymes other than mtAPE have been characterized. In order to identify and characterize mtAPE, we purified the APE activity from beef liver mitochondria to near homogeneity, and showed that the mtAPE which has 3-fold higher specific activity relative to APE1 is derived from the latter with deletion of 33 N-terminal residues which contain the nuclear localization signal. The mtAPE-sized product could be generated by incubating 35S-labeled APE1 with crude mitochondrial extract, but not with cytosolic or nuclear extract, suggesting that cleavage of APE1 by a specific mitochondria-associated N-terminal peptidase is a prerequisite for mitochondrial import. The low abundance of mtAPE, particularly in cultured cells might be the reason for its earlier lack of detection by western analysis.

MeSH Terms
Amino Acid Sequence Animals Cattle DNA-(Apurinic or Apyrimidinic Site) Lyase/genetics,isolation & purification,metabolism Enzyme Stability Humans Kinetics Mice Mitochondria/enzymology Mitochondria, Liver/enzymology Mitochondrial Proteins/genetics,isolation & purification,metabolism Molecular Sequence Data Peptide Hydrolases/metabolism Recombinant Proteins/metabolism Sequence Deletion
Chemicals
Mitochondrial Proteins Recombinant Proteins Peptide Hydrolases APEX1 protein, human DNA-(Apurinic or Apyrimidinic Site) Lyase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Chattopadhyay Ranajoy
Sealy Center for Molecular Science, Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555-1079, USA.
Wiederhold Lee
Szczesny Bartosz
Boldogh Istvan
Hazra Tapas K
Izumi Tadahide
Mitra Sankar
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkl177
Published
2006-00-00
Epub
2006-00-14
Pages
2067-76
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1440881
Subset
IM
Grants
NIA NIH HHS · P01 AG021830 · United States
NCI NIH HHS · R01 CA098664 · United States
NIEHS NIH HHS · ES08457 · United States
NIEHS NIH HHS · R01 ES008457 · United States
NCI NIH HHS · R01 CA053791 · United States
NCI NIH HHS · CA98664 · United States
NIEHS NIH HHS · P30 ES006676 · United States
NIEHS NIH HHS · ES06676 · United States
NCI NIH HHS · R56 CA098664 · United States
NCI NIH HHS · R01 CA53791 · United States
External Links
PubMed source DOI: 10.1093/nar/gkl177 PMC Full Text View journal details
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